Abstract
Background and aims
Abnormal apoptosis may result in the persistence of activated intestinal T-cells in inflammatory bowel disease (IBD). We investigated apoptosis in distinct mucosal compartments, and the expression of Fas/Fas ligand and perforin in the inflamed and non-inflamed intestinal mucosa of patients with IBD.
Methods
Colon specimens from 15 patients with ulcerative colitis (UC) and inflamed and non-inflamed mucosa from 15 patients with Crohn’s disease (CD) were analysed for densities and distribution of apoptotic cells determined by the terminal deoxynucleotidyltransferase-mediated dUDP-biotin nick-end labelling (TUNEL) method. Fas, FasL, and perforin-expressing cells were assessed by immunoperoxidase, and with anti-CD3, anti-CD20 and anti-CD68, by double immunofluorescence with confocal microscopy. Quantitative analysis was performed using a computer-assisted image analyser.
Results
Colonic lamina propria (LP) and epithelium from patients with UC showed higher rates of apoptosis than controls, but no difference was shown regarding patients with CD. In LP, co-expression of Fas was reduced with T-cells in inflamed CD mucosa, and with macrophages in all patients with IBD. No difference was found in the expression of Fas on B-cells. Rates of FasL-expressing cells in LP were higher in IBD than in controls, with no correlation with the rates of apoptosis. Rates of perforin-expressing cells in LP were greater in UC than in controls, and correlated to the rates of apoptosis. No difference was shown regarding the inflamed and non-inflamed CD mucosa. Rates of FasL and perforin-expressing intra-epithelial lymphocytes showed no difference among groups.
Conclusions
Increased expression of FasL in IBD colonic LP not parallelled by Fas on T-cells and macrophages may indicate a reduced susceptibility to the Fas/FasL-mediated apoptosis of lymphoid cells. Expression of perforin is correlated to the tissue damage, and may represent the enhancement of a distinct cytotoxic pathway in UC.
Similar content being viewed by others
References
Meier P, Finch A, Evan G (2000) Apoptosis in development. Nature 407:796–801
Hall PA, Coates PJ, Ansari B, Hopwood D (1994) Regulation of cell number in the mammalian gastrointestinal tract: the importance of apoptosis. J Cell Sci 107:3569–3577
Bu P, Keshavarzian A, Stone DD, Liu J, Le PT, Fisher S, Qiao L (2001) Apoptosis: one of the mechanisms that maintains unresponsiveness of the intestinal mucosal immune system. J Immunol 166:6399–6403
Plas DR, Rathmell JC, Thompson CB (2002) Homeostatic control of lymphocyte survival: potential origins and implications. Nat Immunol 3:515–521
Thompson CB (1995) Apoptosis in the pathogenesis and treatment of disease. Science 267:1456–1462
Fiocchi C (1998) Inflammatory bowel disease: etiology and pathogenesis. Gastroenterology 115:182–205
Bouma G, Strober W (2003) The immunological and genetic basis of inflammatory bowel disease. Nat Rev Immunol 3:521–533
Boirivant M, Pica R, DeMaria R, Testi R, Pallone F, Strober W (1996) Stimulated human lamina propria T cells manifest enhanced Fas-mediated apoptosis. J Clin Invest 98:2616–2622
Boirivant M, Marini M, Di Felice G, Pronio AM, Montesani C, Tersigni R, Strober W (1999) Lamina propria T cells in Crohn’s disease and other gastrointestinal inflammation show defective CD2 pathway-induced apoptosis. Gastroenterology 116:557–565
Ina K, Itoh J, Fukushima K, Kusugami K, Yamaguchi T, Kyokane K, Imada A, Binion DG, Musso A, West GA, Dobrea GM, McCormick TS, Lapetina EG, Levine AD, Ottaway CA, Fiocchi C (1999) Resistance of Crohn’s disease T cells to multiple apoptotic signals is associated with a Bcl-2/Bax mucosal imbalance. J Immunol 63:1081–1090
Itoh J, de La Motte C, Strong SA, Levine AD, Fiocchi C (2001) Decreased Bax expression by mucosal T cells favours resistance to apoptosis in Crohn’s disease. Gut 49:35–41
Merger M, Viney JL, Borojevic R, Steele-Norwood D, Zhou P, Clark DA, Riddell R, Maric R, Podack ER, Croitoru K (2002) Defining the roles of perforin, Fas/FasL, and tumour necrosis factor alpha in T cell induced mucosal damage in the mouse intestine. Gut 51:155–163
Simpson SJ, De Jong YP, Shah SA, Comiskey M, Wang B, Spielman JA, Podack ER, Mizoguchi E, Bhan AK, Terhorst C (1998) Consequences of Fas-ligand and perforin expression by colon T cells in a mouse model of inflammatory bowel disease. Gastroenterology 115:849–855
Harvey RF, Bradshaw JM (1980) A simple index of Crohn’s-disease activity. Lancet 1:514
Truelove SC, Witts LJ (1955) Cortisone in ulcerative colitis; final report on a therapeutic trial. Br Med J 4947:1041–1048
Seldenrijk CA, Morson BC, Meuwissen SG, Schipper NW, Lindeman J, Meijer CJ (1991) Histopathological evaluation of colonic mucosal biopsy specimens in chronic inflammatory bowel disease: diagnostic implications. Gut 32:1514–1520
Truelove SC, Richards WC (1956) Biopsy studies in ulcerative colitis. Br Med J 4979:1315–1318
Watson AJ (1995) Review article: manipulation of cell death—the development of novel strategies for the treatment of gastrointestinal disease. Aliment Pharmacol Ther 9:215–226
Merritt AJ, Potten CS, Watson AJ, Loh DY, Nakayama K, Nakayama K, Hickman JA (1995) Differential expression of bcl-2 in intestinal epithelia. Correlation with attenuation of apoptosis in colonic crypts and the incidence of colonic neoplasia. J Cell Sci 108:2261–2271
Ju ST, Panka DJ, Cui H, Ettinger R, el-Khatib M, Sherr DH, Stanger BZ, Marshak-Rothstein A (1995) Fas(CD95)/FasL interactions required for programmed cell death after T-cell activation. Nature 373:444–448
Suda T, Takahashi T, Golstein P, Nagata S (1993) Molecular cloning and expression of the Fas ligand, a novel member of the tumor necrosis factor family. Cell 75:1169–1178
Brunner T, Mogil RJ, LaFace D, Yoo NJ, Mahboubi A, Echeverri F, Martin SJ, Force WR, Lynch DH, Ware CF, Green DR (1995) Cell-autonomous Fas (CD95)/Fas-ligand interaction mediates activation-induced apoptosis in T-cell hybridomas. Nature 373:441–444
Miyawaki T, Uehara T, Nibu R, Tsuji T, Yachie A, Yonehara S, Taniguchi N (1992) Differential expression of apoptosis-related Fas antigen on lymphocyte subpopulations in human peripheral blood. J Immunol 149:3753–3758
Moller P, Koretz K, Leithauser F, Bruderlein S, Henne C, Ouentmeier A, Krammer PH (1994) Expression of APO-1 (CD95), a member of the NGF/TNF receptor superfamily, in normal and neoplastic colon epithelium. Int J Cancer 57:371–377
Ashany D, Song X, Lacy E, Nikolic-Zugic J, Friedman SM, Elkon KB (1995) Th1 CD4+ lymphocytes delete activated macrophages through the Fas/APO-1 antigen pathway. Proc Natl Acad Sci USA 92:11225–11229
Strater J, Wellisch I, Riedl S, Walczak H, Koretz K, Tandara A, Krammer PH, Moller P (1997) CD95 (APO-1/Fas)-mediated apoptosis in colon epithelial cells: a possible role in ulcerative colitis. Gastroenterology 113:160–167
Ueyama H, Kiyohara T, Sawada N, Isozaki K, Kitamura S, Kondo S, Miyagawa J, Kanayama S, Shinomura Y, Ishikawa H, Ohtani T, Nezu R, Nagata S, Matsuzawa Y (1998) High Fas ligand expression on lymphocytes in lesions of ulcerative colitis. Gut 43:48–55
Jenkins D, Seth R, Kummer JA, Scott BB, Hawkey CJ, Robins RA (2000) Differential levels of granzyme B, regulatory cytokines, and apoptosis in Crohn’s disease and ulcerative colitis at first presentation. J Pathol 190:184–189
Lundqvist C, Melgar S, Yeung MM, Hammarstrom S, Hammarstrom ML (1996) Intraepithelial lymphocytes in human gut have lytic potential and a cytokine profile that suggest T helper 1 and cytotoxic functions. J Immunol 157:1926–1934
Corazza N, Muller S, Brunner T, Kagi D, Mueller C (2000) Differential contribution of Fas- and perforin-mediated mechanisms to the cell-mediated cytotoxic activity of naive and in vivo-primed intestinal intraepithelial lymphocytes. J Immunol 164:398–403
Morimoto Y, Hizuta A, Ding EX, Ishii T, Hongo T, Fujiwara T, Iwagaki H, Tanaka N (1999) Functional expression of Fas and Fas ligand on human intestinal intraepithelial lymphocytes. Clin Exp Immunol 116:84–89
Acknowledgements
This work was supported by grants from the Brazilian Research Council/CNPq, FAPERJ, and Fundação José Bonifácio/FUJB. We thank the technical assistance of Leonardo Andrade from Departamento de Histologia e Embriologia, Universidade Federal do Rio de Janeiro (UFRJ), for the use of the confocal microscopy.
Author information
Authors and Affiliations
Corresponding author
Additional information
Heitor S.P. Souza and Claudio J.A. Tortori contributed equally to this paper
Rights and permissions
About this article
Cite this article
Souza, H.S.P., Tortori, C.J.A., Castelo-Branco, M.T.L. et al. Apoptosis in the intestinal mucosa of patients with inflammatory bowel disease: evidence of altered expression of FasL and perforin cytotoxic pathways. Int J Colorectal Dis 20, 277–286 (2005). https://doi.org/10.1007/s00384-004-0639-8
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00384-004-0639-8